Yeo Christine X, Gilchrist Robert B, Thompson Jeremy G, Lane Michelle
Research Centre for Reproductive Health, Discipline of Obstetrics and Gynaecology, University of Adelaide, Medical School, Adelaide, South Australia 5005, Australia.
Hum Reprod. 2008 Jan;23(1):67-73. doi: 10.1093/humrep/dem140. Epub 2007 Oct 12.
Successful oocyte in vitro maturation (IVM) would eliminate the need for hormonal stimulation used in assisted reproduction techniques. Unfortunately, oocytes matured in vitro have compromised developmental competence possibly due to disrupted oocyte-cumulus communication resulting from inappropriate levels of oocyte-secreted factors such as growth differentiation factor 9 (GDF9). Hence, the aim of this study was to investigate the effects of exogenous GDF9 during IVM of mouse oocytes on development and subsequent fetal viability.
Cumulus-oocyte complexes from pregnant mare's serum gonadotrophin primed mice were cultured with or without 200 ng/ml exogenous recombinant GDF9, 50 mIU/ml FSH and 10 ng/ml epidermal growth factor (EGF). After 18 h, cumulus expansion was scored and oocytes were fertilized in vitro. Cleavage, blastocyst development, blastocyst total, inner cell mass (ICM) and trophectoderm cell numbers were assessed. Viability of embryos was assessed by transfer to recipient females and pregnancy outcome determined at day 15.
Oocytes matured with exogenous GDF9 in the presence of FSH and EGF had higher rates of development, percentage of hatching blastocyst and blastocyst total and ICM cell numbers (all P < 0.05). Although implantation rate and fetal and placental weights were not affected, the number of viable fetuses at day 15 was increased with exogenous GDF9.
Exogenous GDF9 during IVM improved embryo development and fetal viability and provides a promising approach for human IVM.
成功的卵母细胞体外成熟(IVM)将消除辅助生殖技术中使用的激素刺激的必要性。不幸的是,体外成熟的卵母细胞发育能力受损,这可能是由于卵母细胞分泌因子(如生长分化因子9,GDF9)水平不当导致卵母细胞-卵丘细胞通讯中断所致。因此,本研究的目的是探讨外源性GDF9在小鼠卵母细胞IVM过程中对发育及随后胎儿活力的影响。
将来自孕马血清促性腺激素预处理小鼠的卵丘-卵母细胞复合体在含有或不含有200 ng/ml外源性重组GDF9、50 mIU/ml促卵泡激素(FSH)和10 ng/ml表皮生长因子(EGF)的条件下进行培养。18小时后,对卵丘扩展情况进行评分,并将卵母细胞进行体外受精。评估卵裂、囊胚发育、囊胚总数、内细胞团(ICM)和滋养外胚层细胞数量。通过将胚胎移植到受体雌性小鼠体内评估胚胎活力,并在第15天确定妊娠结局。
在FSH和EGF存在的情况下,用外源性GDF9成熟的卵母细胞具有更高的发育率、孵化囊胚百分比、囊胚总数和ICM细胞数量(所有P<0.05)。虽然着床率、胎儿和胎盘重量未受影响,但外源性GDF9使第15天的活胎数量增加。
IVM期间的外源性GDF9改善了胚胎发育和胎儿活力,并为人类IVM提供了一种有前景的方法。